Chemically modified factor ix

Abstract

The present invention discloses a chemically modified FIX, wherein the activation peptide region contains a covalently coupled water-soluble hydrophilic polymer.

1 . Blood coagulation factor IX (FIX) with a FIX activation peptide region (AP region), wherein said AP region comprises a covalently coupled water-soluble hydrophilic polymer, said polymer being absent from biologically produced FIX. 
   
   
       2 . FIX according to  claim 1 , wherein said FIX is recombinant human FIX (rhFIX). 
   
   
       3 . FIX according to  claim 2 , wherein said rhFIX is expressed in CHO- or HEK293-derived cells. 
   
   
       4 . FIX according to  claim 1 , wherein Tyr-155 of FIX is sulfated and/or Ser-158 of FIX is phosphorylated. 
   
   
       5 . FIX according to  claim 1 , wherein said water-soluble hydrophilic polymer is hydroxyethyl starch (HES), polyethylene glycol (PEG), dextran or polysialic acid (PSA). 
   
   
       6 . FIX according to  claim 5 , wherein said water-soluble hydrophilic polymer is attached to FIX via Asn-157 and/or Asn-167 of FIX. 
   
   
       7 . FIX according to  claim 6 , wherein said water-soluble hydrophilic polymer is HES or PSA. 
   
   
       8 . FIX according to  claim 1 , wherein said water-soluble hydrophilic polymer is attached to FIX via Ser-158, Thr-159, Thr-163, Thr-169, Ser-171, Thr-172, Ser-174 or Thr-179, especially via Ser-158, Thr-163, Ser-171 or Ser-174, of FIX. 
   
   
       9 . FIX according to  claim 8 , wherein said water-soluble hydrophilic polymer is PEG. 
   
   
       10 . FIX according to  claim 1 , wherein said water-soluble hydrophilic polymer is attached to FIX via a releasable linker, especially a hydrolysable linker. 
   
   
       11 . Pharmaceutical composition containing a FIX according to  claim 1  and a pharmaceutically acceptable carrier. 
   
   
       12 . The composition according to  claim 11 , wherein FIX has a specific activity of at least 100 international units (IU) of FIX per mg FIX protein, especially at least 200 IU FIX per mg FIX protein. 
   
   
       13 . The composition according to  claim 11 , wherein said pharmaceutically acceptable carrier is an amino acid, preferably L-histidine or glycine, a carbohydrate, preferably sucrose, a tenside, preferably a polysorbate, especially polysorbate 80, or mixtures thereof. 
   
   
       14 . The composition according to  claim 11 , wherein the composition is in lyophilized form. 
   
   
       15 . A method of treating a bleeding disorder, the method comprising the step of administering an effective amount of a pharmaceutical composition according to  claim 11  to a patient in need thereof. 
   
   
       16 . A method according to  claim 15  wherein said bleeding disorder is haemophilia B. 
   
   
       17 . A method for the preparation of a blood coagulation factor IX (FIX) with a FIX activation peptide region (AP region), wherein said AP region comprises a covalently coupled water-soluble hydrophilic polymer according to  claim 1  comprising the following steps:
 providing a FIX molecule comprising a FIX activation peptide region (AP region),   covalently coupling a mixing a water-soluble hydrophilic polymer to said AP region, and   isolating a FIX with a covalently coupled water-soluble hydrophilic polymer in said AP region.   
   
   
       18 . The method according to  claim 17 , wherein said FIX is recombinant human FIX (rhFIX). 
   
   
       19 . The method according to  claim 18 , wherein said rhFIX is expressed in CHO- or HEK293-derived cells. 
   
   
       20 . The method according to  claim 17 , wherein Tyr-155 of FIX is sulfated and/or Ser-158 of FIX is phosphorylated. 
   
   
       21 . The method according to  claim 17 , wherein said water-soluble hydrophilic polymer is hydroxyethyl starch (HES), polyethylene glycol (PEG), dextran or polysialic acid (PSA). 
   
   
       22 . The method according to  claim 17 , wherein said water-soluble hydrophilic polymer is attached to FIX via Asn-157 and/or Asn-167 of FIX. 
   
   
       23 . The method according to  claim 22 , wherein said water-soluble hydrophilic polymer is HES or PSA. 
   
   
       24 . The method according to  claim 17 , wherein said water-soluble hydrophilic polymer is attached to FIX via Ser-158, Thr-159, Thr-163, Thr-169, Ser-171, Thr-172, Ser-174 or Thr-179, especially via Ser-158, Thr-163, Ser-171 or Ser-174, of FIX. 
   
   
       25 . The method according to  claim 24 , wherein said water-soluble hydrophilic polymer is PEG. 
   
   
       26 . The method according to  claim 17 , wherein said water-soluble hydrophilic polymer is attached to FIX via a releasable linker, especially a hydrolysable linker.